Method for Treating Oncological, Virulent and Somatic Diseases, Method for Controlling Treatment Efficiency, Pharmaceutical Agents and Composition for Carrying Out Said Treatment

ABSTRACT

The invention relates to medicine and veterinary science and discloses a novel method for treating oncological, virulent and somatic diseases whose main target for therapeutic action is embodied in the form of DNA which freely circulates in blood plasma (and other liquid media) and originates from tumoral and mutant cells or cells infected by bacteria, fungi or protozoan and from different microorganisms which reside in the organism thereof. Said invention also relates to novel pharmaceutical compositions and to the use thereof for treating oncological diseases and infectious states provoked by bacteria, fungi and protozoa and non-infectious somatic diseases and states produced by accumulation of somatic mutations in cells of organism. Medicinal and immunological compositions, sorption and physico-chemical engineering and the method for using them in order to treat malignant tumors and other diseases are also disclosed.

TECHNICAL FIELD

The invention relates to medicine and veterinary science and discloses anovel method for treating oncological, virulent and somatic diseaseswhen the main target for therapeutic action is embodied in the form ofDNA which freely circulates in blood plasma (and other liquid media) andoriginates from tumoral and mutant cells or cells infected by bacteria,fungi or protozoan and from different microorganisms which reside in theorganism thereof. Said invention also relates to novel pharmaceuticalcompositions and to the use thereof for treating oncological diseasesand infectious states provoked by bacteria, fungi and protozoa andnon-infectious somatic diseases and states produced by accumulation ofsomatic mutations in cells of organism. Medicinal and immunologicalcompositions, sorption and physico-chemical engineering and the methodfor using them in order to treat malignant tumors and to prevent theirrelapses as well as for treatment of infections, atherosclerosis,diabetes and slowing the aging process. Proposed method is based oncompletely new principle of therapeutic action, provide strong antitumorand antimicrobial efficacy and could be used in therapy of oncologicaldiseases, different infections and non-infectious diseases.

BACKGROUND ART

Populations of cancer cells, developing in patient's organism possessextremely high level of genetic plasticity that is much higher than thatin normal cells. Genetic plasticity within cancer cells populationsallows them to generate in course of disease the phenotypes which areresistant to immune and morphogenic control and which has the capacityfor invasion and metastasis and are resistant to antitumor therapies. Itis known, that selection and clonal expansion of tumor cells comprisethe basis of biological and clinical tumor progression. According tothat knowledge, the modern strategy of anticancer therapy is based onprinciples of tumor cell clones destruction in patient organism throughchemotherapy, radiotherapy, immunotherapy, surgical elimination and byuse of their combinations. Fundamental particularity of all thesemethods is that their sole therapeutic target is a tumor cell. The longterm experience from clinical application of such therapy has shown thatdue to high genetic placticity the most malignant cells becomeinsensitive to the specific therapy used before malignant neoplasm couldbe completely destroyed by said therapy.

There is a great medical need for new antitumor drugs less toxic thanexisting ones or can potentiate thre efficacy of existing methods. Aswell it is a need for new antitumor drugs that can decrease toxicity ofcurrently available methods without decreasing of their efficacy.

The circulation of DNA molecules in blood plasma of cancer patients andhealthy people was known from some publications (P. Anker et al.,Clinica Chimica Acta, v. 313, 2001, pp. 143-146; Fedorov N. A. et. al.,Bull. Exp. Biol. Med., v102,1986, pp 281-283. Patent (U.S. Pat. No.5,952,170) disclose the determination of DNA content in blood plasma fordiagnostic and prognostic purposes in course of cancer disease. Patents(U.S. Pat. No. 6,465,177 and U.S. Pat. No. 6,156,504) disclose the useof blood plasma DNA for mutation's determination in oncogenes andmicrosatellite gene regions, for genetic instability studying in tumorsand the use of these results for diagnostic, monitoring and prognosticpurposes in the course of disease.

Sugihara S. et al. (1990, 1993) studied

chemotrypsine and desoxiribonuclease I (DNase I) enzymes influence onautologic and heterologic adhesion of tumor cells during theirmetastasing. They have shown that systemic introduction of DNase I couldslow metastases development. The effect they find was therapeuticallyinsufficient. Authors make conclusion that DNase I can be used togetherwith surgical elimination of tumor for prevention of tumor cells'hematogenic dissemination. Authors meant the influence of DNase I ontumor cells' cytoplasmatic membrane but not on destruction of freecirculating DNA. Accordingly, the regime and doses that were used couldnot cause the decrease of circulating DNA level.

Torchilin V. P. (2001), Patent (U.S. Pat. No. 5,780,033), disclose thetherapeutic use of autoantibodies that are able to bind with cytoplasmicand nuclear membranes of tumor cells and with protein-DNA complex comingfrom dead tumor cells. It is clear from the text of application that thedisclosure relates solely to antibodies against protein antigenicdeterminants. In our case anti-DNA antibodies and anti-DNA abzymes areused. Moreover, therapy disclosed by authors was trended againstphagocytosis of nucleosomes on the surface of tumor cells and thatexclude the possibility of therapeutic regimes sufficient for bindingand removal of circulating blood plasma DNA.

There is practically no data concerning circulation of bacterial DNA inblood. In human organism all microorganisms exist in biofilm communities(Davey M. E. Otoole G. A. 2000. Microbial biofilms: from ecology toMolecular ganetics. Microbiol. Mol. Genet. 64:847-867). Biofilms areformed by bacterial cells, associated with extracellular matrix (Tez V.V. 1999. Formation and structure of mixed bacterial communities. APMIS,107:645-654). In biofilm matrix we have found extracellular DNA thatpenetrates from living cells. Our data also indicates that bacterial DNApresent in blood plasma of infected human and its quantities andcomposition could change subject to development of certain infections.It is also known that DNA can get into the outer environment as a resultof cells death, for example in the focus of inflammation. DNA as polymerincreases biofluids viscosity and that negatively influence the courseof the disease, preventing elimination of toxins, pathogens and necroticdebris. It is known that-DNase (Gentech-Roch) “Pulmozyme” is used forCystic Fibrosis treatment by inhalation. The therapeutic action is basedon dilution of broncho-alveolar secret, and does not relate to breach ofgenetic information traffic.

There is no human or animal data, regarding the genetic composition ofDNA, circulating in blood plasma. There is no data regarding bloodplasma DNA analysis avoiding PCR (Polymerase Chain Reaction). The use ofPCR can considerably distort the representation of DNA compositionbecause of amplification primer's specificity. So far, the geneticanalysis of plasma DNA in cancer patients were done by PSR method orblot hybridization and was directed to studying of genome changes incertain specific genome areas (for example in microsatellites andspecific genes). (Sanchez-Cespedes M., et al., Ann Oncol, 1998, v9(1),pp113-116; Sozzi G., et al., Clin Can Res, 1999, v5 (10), pp 2689-2692;Chen X. Q., et al., Nat Med, 1996, v2(9), pp 1033-1035).

Thus, there is no data available about genetic repertoire of DNA whichcirculates in blood plasma of patients with oncopathology, infections,somatic pathology and healthy people, about the biological role of ofblood plasma circulating DNA and about possible therapeutic effect ofblood plasma circulating DNA destruction or inactivation in thesediseases treatment.

DISCLOSURE OF THE INVENTION

As a result of our work upon the invention we have unexpectedly foundthat blood plasma circulating DNA from oncological patients containunique quantitative and qualitative repertoire of genes and regulatorygenetic elements, which is strongly different from that of human genome.Circulating blood plasma DNA of oncological patients for the most partcontains unique human genes, including genes that are associated withmaintenance and formation of malignancy.

It was shown that circulating blood plasma DNA of patients withoncopathology participates in intercellular transport of geneticinformation within tumor cells' population in patient organism. Currentinvention disclosure methods of free blood plasma circulating DNAdestruction and inactivation which lead to suppression of tumordevelopment in host organism. The invention also disclosure new methodfor identification of genomic structures which are involved in tumorprogression and human genome operation. This aspect of invention isconnected with DNA samples isolation from blood plasma of healthy peopleand patients with oncological diseases, cloning and sequencing them.

It was found out that DNA from biofilm matrix secreted by differentbacteria enters into the blood and biological fluids of human andanimals. It was stated that the presence of extracellular DNA is one ofconditions necessary for microbial infection development.

Invention includes destruction or inactivation of circulating microbialDNA as method for treatment and prophylaxis of diseases caused by them.

It was also found out that DNA circulating in blood of healthy peopleplay role in somatic mosaicism's development. Binding, destruction orinactivation of that DNA could suppress the somatic mosaicismdevelopment. Binding, destruction or inactivation of circulating bloodplasma DNA could be of therapeutic value in treatment of diseases whichprogression is linked with somatic mosaicism development.

One aspect of invention discloses the pharmaceutical compositions andnon pharmaceutical methods of free circulating DNA destruction orinactivation in blood plasma of patients with oncological or infectiousdiseases.

Another invention aspect discloses treatment method of patients' withoncopathology, infections, somatic diseases and method of extension oflife time, which are based on administration of pharmaceutical compoundsor application of non pharmaceutical methods providing the destructionor inactivation to free circulating blood plasma DNA.

Another aspect of invention provide the way of treatment efficacycontrol, when said treatment is directed to destruction or inactivationof free circulating blood plasma DNA, then the monitoring of freecirculating blood plasma DNA and determination of tumor-specific geneticmarkers or genetic microbial markers presence in blood plasma could beused for treatment efficacy control.

Methods of patients' treatment with oncopathology and infectiousdiseases based on pharmaceutical compounds administration or use of nonpharmaceutical methods that lead to destruction or inactivation of freecirculating blood plasma DNA, when such treatment is combined with useof antitumor or antimicrobial therapy were disclosed.

Genetic placticity of tumor cells, which allows them to accumulate andmaintain signs that comprise “malignant phenotype”, reveals itself ongenetic and chromosomal levels by loss, acquiring or changing of DNAconsequences, starting from single nucleoside up to whole chromosomes(Loeb K. R. et. al., Carcinogenesis, v21, 2000, pp. 379-385).

Source of such variability is thought to be a special modus operandy oftumor cells' genetic apparatus. It is characterized by increasedfrequency of spontaneous mutations while activity of reparative systemis decreased and control system of genetic homeostasis is switched off(Schmutte C., et al., Anticancer Res., 1999, v 19, pp. 4665-4696).

It is thought that “mutator” phenotype of tumor cells (Loeb L. A.,Cancer Research, 2001, v61, pp. 3230-3239), dynamic heterogeneity withintumor cell clones (Heppner G. H. et al., International Review ofCytology, 1998, v177, pp. 1-56) and numerous rounds of selection in thecourse of progression (Cahill D. P. et al., Trends in Cell Biology, v9,pp. M57-M60 ; Rubin H., Adv Cancer Res, 2001, v83, pp. 159-207; P.Nowell, Seminars in Cancer Biology, v 12, 2002, pp. 261-266) lead toselection and following expansion of the most malignant tumor clones.According to this knowledge the modern methods of malignant neoplasm'streatment are based on principle of tumor cells destruction in patientorganism.

In the process of investigation we have unexpectedly found thataccumulation of genetic changes that are essential for “malignantphenotype” formation in clinically advanced tumor is a result ofcooperative interclonal interaction inside population of tumor cells inpatients' organism, based on horizontal transfer of genetic information

Free circulating DNA in blood plasma of oncological patients is amessenger of such cooperative relations. It realizes intrapopulationalinterclonal transport of genes, which are responsible for “malignantphenotype” formation.

Destruction or inactivation of free blood plasma circulating DNA willlead to inability of tumor cells to maintain necessary level of geneticvariability in organism and loss ability to maintain “malignantphenotype” (growth, metastasing, insensitivity to therapy). Suchtherapeutic intervention has its own therapeutic value as well as itincreases efficacy of traditional therapeutic methods.

BRIEF DESCRIPTION OF THE DRAWINGS

P-glycoprotein expression in murine tumors

FIG. 1 Results of immunochemical staining of mice tumor's histologicalslices after administration of 5 day course of Doxorubicin therapy (2mg/kg every day intravenous) and DNase 1 (0.5 mg/kg four times a dayduring 5 days).

-   -   A—Doxorubicin+DNase    -   B—Doxorubicin

PREFERRED EMBODIMENT

Isolation of free circulating DNA from blood plasma.

Fresh blood plasma (not more than 3-4 hours after isolation) withanticoagulant (sodium citrate) addition was centrifuged onFicoll-PlaquePlus (Amersham-Pharmacia) step at 1500 g for 20 minutes atroom temperature. Plasma (½ of all amount) was neatly isolated avoidingtouching cells sediment on ficoll step and was centrifuged at 10 000 gfor 30 minutes to eliminate cells and debris. Supernatant was taken awaynot touching sediment and up to 1% of sarcosile, up to 50 mM of tris HClpH 7,6, up to 20 mM EDTA, up to 400 mM NaCl and equal volume ofphenol—chloroform mixture 1:1 were added. Received emulsion wasincubated at 65° C. for 2 hours than phenol-chloroform was separated bycentrifuging at 5000 g during 20 minutes at room temperature.Deproteinization by phenol-chloroform method was identically repeatedfor three times after what water phase was processed by chloroform andafter it by diethyl ether.

Organic solvents' separation was done by centrifuging at 5000 g during15 minutes. Equal part of isopropanol was added to water phase andincubated during night at 0° C.

After sedimentation nucleic acids were separated by centrifuging at 10000 g during 30 minutes. Sediment of nucleic acids was dissolved inbuffer that consisted of 10 mM tris—HCl, pH 7,6, 5 mM EDTA and wasinflicted on step made from chlorine cesium gradient (1M, 2.5M 5.7) incentrifuge test tube for SW60Ti rotor. DNA volume was 2 ml, volume ofeach CsCl step was 1 ml. Ultracentrifuging was done in L80-80 (Beckman)centrifuge for 3 hours at 250000 g. DNA was isolated according tofractions from the step's surface 5.7M. Fractions were dialized during12 hours. mM tris-HCl, pH 7,6, 1 mM EDTA at 4° C. will be added. DNApresence in fractions was defined by agarose electrophoresis with DNAvisualization by ethidium bromide. DNA amount was spectrophotometricallyestimated (Beckman DU 470) in cuvette with volume 100 mkl, using 220-320nm spectrum. Average runout of DNA was 10-20 ng according to 1 ml ofplasma.

Cloning and sequencing of blood plasma DNA.

We have developed new method of DNA isolation and cloning from bloodplasma, that allows to construct not amplified plasmide library of suchDNA with representativeness up to million clones with average size300-500 base pair isolated from 50 ml of blood, even taking into accountsignificant amount of elevated liposaccharides level and non identifiedmixtures that troubled purification of nucleic acids. So representativeanalysis can be done with less amount of plasma pattern—10-20 mldepending on pathological contaminates' presence.

Isolated according to above-mentioned method DNA was deproteinized withthe use of proteinase K (Sigma) at 65° C. for tightly-bound proteinselimination. After deproteinization DNA was processed byphenol-chloroform at 65° C. and sedimented by 2.5 volumes of ethanolduring night. After it DNA was processed by EcoRI restrictase during 3hours or by Pfu polymerase (Stratagene) at the presence of 300 mkM ofall desoxynucleothydethreephosphates for “sticky” edges elimination.Completed DNA was phosphorylated by polynucleotide kinase T4 (30U, 2hours). Received samples/preparations were ligated in Bluescript(Stratagene), plasmid digested by EcoR1 or PvuII accordingly anddephosphorylated by alkaline phosphatase CIP (Fermentas) during 1 hour.1 mkg of vector and 0.1-0.5 mkg of serum DNA were usually used forligation. Ligation was done by Rapid Ligation Kit (Roche) use for 10hours at 16° C. Volume of ligase mixture was 50 mkl. Ligated library wastransformed into DH12S (Life Technologies) cells with electroporator(BioRad) use. For transformation of one library 12-20 electroporationcuvettes were used. Dilutions of the library at concentrations 10⁻⁴,10⁻⁵ and 10⁻⁶ were plated for control on dishes with 1.5% agar and LBmedia, supplemented with 100 mkg\ml ampicillin. In both cases library'srepresentativeness was approximately 2-3×10⁶ clones.

Theoretically set of DNA sequences that circulate in plasma shouldcorrespond to set of genome's DNA sequences. Usually cells apoptosis isaccompanied by quantitative and nonspecific DNA degradation before itsexit out of the cell, so the most wide spread DNA in plasma should berepetitive elements of genome in proportion that correspond tononspecific degradation of DNA.

Such elements are L1 repeats, satellite DNA, Alu, MER, MIR, THE repeatsand some others. Quantity of unique sequences should be small inaccordance to their small percent in human genome they may be notdetected in cloning DNA without PCR.

Blood plasma DNA library of oncological patient with clinically advancedtumor stage.

We have constructed blood plasma DNA library of patient with diagnosedadvanced stage mesothelioma. Representativeness of library was 8.5×10⁵clones, that is a good result, taking into account rather small amountof DNA (5 □g) received after purification from non character for healthydonors liposaccharides that were in extremely high concentrations atplasma of patient.

We have got the unexpected results after analysis of 96 clones withlength from 300 up to 1000 base pairs. (It is necessary to mention thatonly one clone was not identified as human DNA. For all otherscorrespondent information from HumanGenBank that identifies DNA of theseclones as human DNA was received.) As mentioned above, according to datafrom literature it is logically to assume that there will be a lot ofhighly repetitive elements in DNA samples.

But at least 55 out of 96 clones presented unique sequences of humanDNA. Taking into account real ratio of repetitive and unique elements ofhuman genome (95% to 5%) it is obvious that blood plasma DNA repertoireof this patient differs a lot from human genomic DNA repertoire. In thissample an abrupt enrichment by unique DNA sequences is observed.

For 15 out of 55 unique DNA fragments that were identified duringsequencing of 96 clones from the library of blood plasma DNA, functionsor product of correspondent gene were identified. Tables 1-15 presentlist of these sequences and information about their participation information and maintenance of “malignant phenotype”. TABLE 1Participation in Clone Product oncogenesis Source Clone Member of GPlaying major role in Steeg P. S., Nat 24 protein - coupled cancer cellsignalling. Rew receptor family Linked with cell Cancer, 2003, v. 3, pp.transformation, 55-63. supression of Raj G. V., J apoptosis, hormoneUrology, 2002, independence and v. 167, pp. 1458- metastasis 1463. HoffA. O., Neoplasia, 1999 v. 1, pp. 485-491.

TABLE 2 Participation in Clone Product oncogenesis Source CloneSnf2-coupled Transription activator. Thaete C., Hum 43 CBP activatorFamily members Mol protein (SCRAP) linked with synovial Genet, 1999, v.8, pp. sarcoma and 585-91. leukaemia Monroy M, A., J development BiolChem,. 2001, v. 276, pp. 40721-40726 Lee D. W., Cancer Res., 2000, v.60, pp. 3612- 3622.

TABLE 3 Participation in Clone Product oncogenesis Source Clone SRY-boxTranscription Graham J. D., J 51 containing gene modulator. ExpressedMol endocrinol, during embryogenesis. 1999, v. 22, pp. 295- Familymembers 304. linked with Lee C. J., J medulloblastomas, Neurooncol,gonadal tumors, highly 2002, v. 57, pp. 201- metastatic melanoma. 214.Uehara S., Cancer Genet Cytogenet, 1999, v. 113., pp. 78-84. Tani M.,Genomics, 1997, v. 39, pp. 30- 37

TABLE 4 Participation in Clone Product oncogenesis Source CloneProtein-tyrosine Family members Hunter T., Philos 72 kinase playingmajor role in Trans R Soc Lond cancer. Some PTK are B Biol Sci, cellularspecific 1998, v. 353., pp. oncogenes products. 583-605. Scheijen B.,Oncogene, 2002, v. 21., pp. 3314-3333.

TABLE 5 Participation in Clone Product oncogenesis Source CloneFibroblast Family members Chen W. T, 83 activation protein playing rolein cancer Enzyme alpha; cell surface invasion and Protein, 1996, v. 49.,serine protease metastasis. The pp. 59-71. product is active in ScanlanM. J., Proc cancer stroma and Nat Acad Sci USA, different carcinomas.1994, v. 91, pp. 5657-5661. Mathew S., Genomics, 1995, v. 25, pp. 335-337.

TABLE 6 Participation in Clone Product oncogenesis Source Clone Braintestican Proteoglycan with Genini M., 86 unknown function. Int J Cancer,Linked with neoplastic 1996, v. 66, pp. 571- phenotype of 577. embryonalrhabdomyosarcoma cells.

TABLE 7 Participation in Clone Product oncogenesis Source Clone KRABdomain, Family members are Oguri T., 152 Zn-finger proteins known astranscription Gene, 1998, v. 222, repressors. Linked pp. 61-67 withearly Gou D. M., Biochim embryogenesis, Biophys Acta, neuroblastoma,Ewing 2001, v. 1518, pp. sarcoma, 306-310 Tcell lymphoma, in Margolin J.F., Proc progression and Nat Acad Sci USA, chemoresistance in 1994,v/91, pp. 4509- lung cancer. 4513. Bellefroid E. J., EMBO J, 1993, v.12, pp. 1363-1374 Gonzales-Lamuno D., Pediatr Pathol Mol Med, 2002, v.21, pp. 531-540. Marilee J. W., Gene, 1994, v. 152, pp. 227-232.

TABLE 8 Participation in Clone Product oncogenesis Source Clone Antigenlinked Antigen recognized by J. Immunol. 166(4), 190 with melanomaautologous tumor 2871-2877, 2001 infiltrating lymphocytes.

TABLE 9 Participation in Clone Product oncogenesis Source CloneN-cadherin Cell adhesion Hazan R. B., J Cell 167 molecule with majorBiol, role in cancer growth, 2000, v. 148, pp. invasion and 779-790.metastasis. Li G., Cancer Res, 2001, v. 61, pp. 3819-3825. Tran N. L., JBiol Chem, 2002, v. 277, pp. 32905-32914.

TABLE 10 Participation in Clone Product oncogenesis Source Clone FAF1:Fas Phosphoprotein known Jensen H. H., Int J 197 associated factor to bethe proapoptosis Biochem Cell Biol, 1 factor. 2001, v. 33, pp. 577- 589.Ryu S. W., Biochem Biophys Res Commun,

TABLE 11 Participation in Clone Product oncogenesis Source CloneInterleukin 7 Proposed as essential Trinder P., Int J 114paracrine\autocrine Oncol, growth factor for 1999, v. 14, pp. 23-variety of cancers. 31. Cosenza L., Cell Signalling, 2002, v. 14, pp.317- 325.

TABLE 12 Participation in Clone Product oncogenesis Source Clone DEADBox RNA Family members Iggo R. D., Mol Cell 208 helicase-like involvedto RNA Biol, protein methabolism. Linked 1991, v. 11, pp. 1326- toexponential cell 1333. growth in cancer. Causevic M., Oncogene, 2001, v.20, pp. 7734- 7743.

TABLE 13 Participation in Clone Product oncogenesis Source Clone Lipin 1One of tumor cells' Brachat A. et. al., 97 response regulators onOncogene, cytotoxic compounds. 2002, v. 21, pp. 8361- 8371

TABLE 14 Participation in Clone Product oncogenesis Source Clone DyneinTakes part in transport Bull J. H., et. al., Br J 121 of p53 protein, isCancer, 2001, v. 84, pp. hypersecreted at 1512-1519. cancer of prostateand Giannakakou P., hepatocellular cancer. et. al., Nat Cell Biol, 2000,v. 2, pp. 709- 717 Jiang J., et. al., Gene, 2001, v. 281, pp. 103- 113.

TABLE 15 Participation in Clone Product oncogenesis Source Clone Rampprotein Linked with human Cheung W. M., 178 embrional carcinoma et. al.,cells' development. J Biol Chem, 2001, v. 276, pp. 17083- 17091

In this way 14 out of 15 sequences with identified function or proteinthat encodes different products (protein kinases, growth factors,proteinases, adhesive molecules and regulatory nuclear proteins) aredescribed in literature as related to “malignant phenotype” formationand maintenance. Only product of 197^(th) clone identified aspro-apoptotic factor is not clearly linked with malignant progression.Though there is data concerning relationship between high apoptoticactivity of tumor with its progression (Nishimura R., et al., J SurgOncol, 1999, v. 71, pp. 226-234) and possible role of apoptoticinductors in formation and maintenance of immunosupression in malignantgrowth (O'Connel J., et al., Dis Esophagus, 1999, v. 12, pp. 83-89).

The most significant presence from repetitive elements in this materialwas alpha-satellite DNA (30 clones). It is possible to say that thatalpha-satellite DNA was the only highly repeated element from humangenome, which behaves exactly as repeat in this material. The rest ofhighly repetitive elements were presented in material as one or severalclones (L1 variant and MLT26), or were not found among patterns (MER,Alu, THE, MIR, {tilde over (□)}satellite). Based on today's knowledgeone can assume that plasma blood composition for the most part shouldrepeat composition of genome DNA, so listed repeats should berepresented in the major part of clones while unique and moderatelyrepeated consequences in analysis of such a small number of clonesshould not be recognized at all. Received result clearly indicates onspecial way of plasma DNA formation in oncological patients. Anotherunexpected result is that of finding in the material of two newmoderately repeated sequences—duplicones, that were recently unknown,also support the evidence of special way of plasma DNA formation inpatients with malignant tumor. For the first time duplicones were foundin human genome less then two years ago. Known duplicones (Eichler E.E., et al., Genome Res, 1998, v8, pp. 791-808; Ji Y., et al., GenomeRes, 2000, v. 10, pp. 597-610; Pujana M. A., et al., Genome Res, 2001,v. 11, pp. 98-111) are extensive regions of DNA that were multiplied forseveral times in the frame of one chromosome (unlike other repeats thatare randomly allocated in genome). Duplicones' formation and expansionis connected with different genetic syndromes (for examplePrader-Willi/Angelmane syndrome), with multigenic families' evolutionsuch as MHS (Shiina T., et al., Proc Nat Acad Sci USA, 1999, v. 96, pp.13282-13287) and with chromosome instability in tumors.

It is necessary to mention that analysis of clones received from bloodplasma DNA of patient has given us unexpected results.

Blood plasma DNA of oncological patient is highly enriched with uniquegenes. 55 out of 96 analyzed clones contain fragments of genome's uniquesequences. 14 out of 15 sequences with identified in functions refer toprocess of tumor progression and maintenance of “malignant phenotype”.

Strong impoverishment of the most wide spread human repeat such as MER,Alu, THE, MIR, {tilde over (□)}satellites is found in plasma DNAmaterial.

Finding of two consequences with previously unknown duplicones'characteristics indicates on duplicones' representativeness in such DNAsamples.

DNA library of healthy donor's blood plasma.

For method's value proof of blood plasma DNA cloning and sequencing foridentification of genome's unique genetic consequences we haveconstructed DNA library of healthy donor's blood plasma. It is knownthat plasma of clinically healthy people also contains DNA but insignificantly less amount than plasma of oncological patients (ShapiroB., et al., Cancer, 1983, v. 51, pp. 2116-2120).

Representativeness of library was near 8×10⁵ clones. We have gotinteresting result after analysis of 70 clones with length from 300 upto 1000 base pairs. We found out that 58 out of 70 analyzed clones areunique DNA sequences of human genome. After searching the HumanGenBank,we identified the function or product of correspondent gene for 14 outof 58 unique DNA fragments.

Only 12 clones contained fragments of repetitive sequences, herewithwithout of alpha-satellite DNA dominance.

So, it was unexpectedly found out that blood plasma DNA of healthypeople and oncological patients for the most part contain uniquefragments of human genome. In the case of oncological pathology uniquesequences of blood plasma DNA correspond to genes which products takepart in the formation and maintenance of tumor cell's “malignantphenotype”.

Basing on this unexpected discovery we have suggested that DNAcirculating in patient's blood can be messenger of horizontal geneticinformation transfer during the course of oncological diseases,assisting to accumulation and spreading of genes that are necessary for“malignant phenotype” formation and maintenance within population oftumor cells.

Somatic mosaicism is a condition that is a result of geneticallynon-identical somatic cells' presence in organism. Modern visionpresents that many non tumor and noninfectious (so called somatic) humandiseases (for example atherosclerosis, diabetes, nonspecific chroniclung diseases and so on), including aging process, are connected withappearance and spreading (expansion) in the process of individualdevelopment of somatic cells' clones that have mutant genes.(Youssoufian H., et al., Nature Rew. Genet., 2002, v. 3, pp. 748-758; J.Vijg, Mutation Res., 2000, v. 447, pp. 117-135; R. Erickson, MutationRes., 2003, v. 543, pp. 125-136; Andreassi M., Mutation Res., 2003, v.543, pp. 67-87; Anderson G., et al., Trends in Pharmacological Sci.,2003., v. 24, pp. 71-76).

Bright example of such process is progression of mitochondrialheteroplasmia (expansion of mutant mitochondrial DNA) at differentdiseases and in the aging process (E. Jazin et al., Proc Nat Acad SciUSA, 1996, v. 93, pp. 12382-12387; Michikawa Y. et al., Science, 1999,v. 286, pp. 774-779; Calloway C. et al., Am J Hum Gen, 2000, v. 66, pp.1384-1397).

There are two alternative models of somatic mosaicism's appearance. Thefirst is appearance of somatic mosaicism as a result of numerous “denovo” mutations in polyclonal cellular pool. The second model is clonalexpansion of mutant cells' clone (Khrapko K., et al., Muation Res.,2003, v. 522, pp. 13-19).

In the process of work above the invention we have found that DNAcirculating in the blood of healthy people play significant role insomatic mosaicism's development and its binding, destruction orinactivation inhibits development of somatic mosaicism. Binding,destruction or inactivation of circulating in blood plasma DNA providestreatment effect at diseases which appearance is connected with somaticmosaicism's development.

Examples that are mentioned later indicates role of circulating in bloodof oncological patients DNA in the development of tumor's resistance tochemotherapy, development of metastasing process, in sepsis developmentand in some other pathological conditions. High therapeutic effect ofblood plasma DNA's binding, destruction or inactivation is found.

Role of Free Circulating DNA in Tumor Progression

Materials and Methods.

Bovine pancreatic DNase I (Fermentas) and human recombinant DNase I(Dornase; Gentech) were used. DNase I solution for injections wereprepared by dissolving of DNase matrix solution in sterile phosphatebuffer directly before administration. Cyclophosphamide and Doxorubicinwere dissolved according to instruction.

In series of in vitro experiments we have not seen any suppressiveeffect of DNase I on tumor cells' growth (DNase I concentration was upto 100 mkg/ml).

Blood plasma DNA of tumor-bearing mice was received according totechnique described above. Highly metastatic and low metastatic stainsof mice' Luice lung carcinoma and Erlich carcinoma were used. Cells werecultivated in RPMI-1640 media with adding of 10% embryonic calf serum,1% penicillin—streptomycin in 5% carbon dioxide atmosphere. For tumorinduction in mice, cells were cultivated until the monolayer, than weredetached with trypsin-EDTA solution. Cells were washed trice bycentrifuging in phosphate buffer and were resuspensed until theirconcentration have become 0.5×10 ⁷/ml. Viability was defined byinclusion of methylene blue in hemocytometer. Suspensions that contained95% of viable cells were used for administration to animals.

Mice of C57B1 line and white not thoroughbred mice from “Rappolovo”animal house were used. Weight of animals was 24-26 grams. Animals werecontained in cages, for 6-7 mice per one cage on a standard diet withoutwater restriction. LLC cells in dose of 5×10⁵ in 100 mkl of phosphatebuffer were injected in soft tissues of thigh. Erlich tumor wassubcutaneously replanted on the right side by injection of 0.2 ml 10%cells' suspension in isotonic sodium chloride solution.

In some experiments DNA presence in blood plasma of mice was studied.

DNA was isolated by method that is mentioned above. DNA amount wasspectrophotometrically measured.

The invention is illustrated by following examples.

EXAMPLE 1

Influence of DNase I Administrated Twice a Day on Erlich Tumor Growth atMice

Group 1—10 mice transplanted with Erlich carcinoma (control).

Group 2—10 mice transplanted with Erlich carcinoma. Mice wereintraperitoneally injected with DNase at 1 mg/kg dose in 200 ml ofphosphate buffer twice a day starting from 3 up to 7 day after tumortransplantation.

Group 3—10 mice transplanted with Erlich carcinoma. Mice wereintraperitoneally injected with DNase at 2 mg/kg dose in 200 ml ofphosphate buffer twice a day starting from 3 up to 7 day after tumortransplantation.

Results of experiments were estimated according to tumor's growthinhibition (TGI) that were expressed in percents to control datareceived at last day of DNase injection with standard formula using.

Tumor's size on the 7 day after transplantation. Tumor's Group volumeTGI % P 1  86+/−12 — — 2 33 +/− 6 61% p < 0.001 3 34 +/− 7 60% p < 0.001

The data obtained indicates that tumor's growth is significantlyinhibited by DNase injection.

EXAMPLE 2 Inhibition of Erlich Tumor Growth by Use of Different Schemesof DNase Administration

In our experiments DNA circulating in blood plasma of patients was thetherapeutic target of DNase. Prolonged presence of DNase in blood plasmaat catalytically effective concentrations is essential for provision ofmaximal therapeutic effect. According to that DNase myltiplyadministration should provide better therapeutic effect than the sametotal daily dose, administrated just as only two daily injections.

Group 1—10 mice transplanted with Erlich carcinoma (control).

Group 2—10 mice transplanted with Erlich carcinoma. Mice wereintraperitoneally injected with DNase at 1 mg/kg dose in 200 ml ofphosphate buffer twice a day starting from 3 up to 7 day after tumortransplantation.

Group 3—10 mice transplanted with Erlich carcinoma. Mice wereintraperitoneally injected with DNase at 0.5 mg/kg dose in 200 ml ofphosphate buffer four times a day starting from 3 up to 7 day aftertumor transplantation.

Results of experiments were estimated according to tumor's growthinhibition (TGI) that were expressed in percents to control datareceived at last day of DNase injection with standard formula using

Tumor's size on the 7 day after transplantation. Group Tumor's volume T%P 1  98+/−14 — — 3 23 +/− 6 76% P < 0.001 2 32 +/− 6 67% P < 0.001

Received data indicates that tumor's growth is more inhibited byfractional (four times a day) administration of DNase at the same totaldaily dose than twice a day administration. Blood plasma DNAconcentration of mice from 3 group after end of treatment course was38.5 ng/ml, mice from control group had 104.8 ng/ml and mice from the 2group—55.1 ng/ml. Blood plasma DNA concentration of healthy mice was12.0 ng/ml (p<0.01). So, multiply daily administrations of DNase lead tomore significant decrease of blood plasma DNA level and greater tumor'sgrowth inhibition in comparison with twice a day administration of thesame total dose.

EXAMPLE 3 Combined Use of DNase and Antitumor Doxorubicin Compound

Group 1—10 mice transplanted with Erlich carcinoma (control).

Group 2—10 mice replanted with Erlich carcinoma. Mice were intravenouslyinjected with Doxorubicin at 2 mg/kg dose once a day starting from 3 upto 7 day after tumor transplantation.

Group 3—10 mice replanted with Erlich carcinoma. Mice were intravenouslyinjected with Doxorubicin at 2 mg/kg dose once a day starting from 3 upto 7 day after tumor transplantation. As well mice wereintraperitoneally injected with DNase at 0.5 mg/kg dose in 200 ml ofphosphate buffer four times a day starting from 3 up to 7 day aftertumor transplantation.

Group 4—10 mice replanted with Erlich carcinoma. Mice wereintraperitoneally injected with DNase at 0.5 mg/kg dose in 200 ml ofphosphate buffer four times a day starting from 3 up to 7 day aftertumor transplantation.

Results of experiments were estimated according to tumor's growthinhibition (TGI) that were expressed in percents to control datareceived at last day of DNase injection with standard formula using.

Tumor's size on the 7^(th) day after transplantation. Group Tumor'svolume T% P 1 98 +/− 14 — — 2 57 +/− 10 42 P < 0.05 4 20 +/− 6  78 P <0.01 3 0 100%

Received data indicates that tumor's growth is less inhibited byDoxorubicin than by DNase. Combined use of Doxorubicin and DNase presentsynergism in their action leading to complete inhibition of tumor'sgrowth (tumors were not found in any experimental animals).

EXAMPLE 4 Influence of DNase on Bacterial Biofilm Formation

For estimation of possible mechanisms of DNase action on bacteria'interactions with host's organism we have estimated its influence onbiofilm formation.

Experiments were performed on biofilm models, received at laboratorycultivation on the surface of glass. Non-relative Gram-positive(Staphylococcus aureus VT-209) and Gram-negative (Escherichia coliATCC25922) bacteria were used.

Bacteria were sowed at 10⁹ bacteria/ml concentrations and were incubatedat 37° C. during 72 hours on synthetic M9 media that contains additivesnecessary for used bacteria strains. DNase was added in amount of 100mkg/ml right after bacteria' inoculation. Identical volume of phosphatebuffer was added to control. Selective vials were taken for analysisevery 24 hours.

Glasses were washed by PBS buffer, fixed, stained with methylene blueand studied with the use of light microscopy.

As a result it was found out that in DNase presence normal biofilm wasnot formed. Only adhesion of single microorganisms to the glass thathave not lead to the formation of microcolonies and biofilms wasobserved.

Control sowings from vials for definition of colony-forming units (CFU)have shown that at DNase presence there is no death of cells leading todecrease of CFU's numbers in comparison with controls. Received dataindicates that DNase do not cause death of Staphylococci andEscherichia, but prevent their cooperative behavior that leads tobiofilm formation.

EXAMPLE 5 DNase' Use for Treatment of the Experimental Sepsis

White not thoroughbred mice were used for experiment. Animals' weightwas 24-26 grams. Pathogenic strain of Staphylococcus aureus VT-2003R at1×10¹⁰ bacteria/animal dose was inoculated to retroorbital sinus ofanimals. DNase was intraperitoneally administered. Isotonic solution ofsodium chloride or Penicillin was administrated to control groups. Eachexperimental group contained 10 mice. Intraperitoneal administration ofcompound (DNAase) four times a day at 500 mkg/kg dose continued for 1-3days after infection. Penicillin was intramusculary administrated.Efficiency of action was estimated by number of animals that stayedalive after the death of the last one in control group. All animals havedied to the third day of the experiment in control group. Six animalsstayed alive to the third day of the experiment in the group thatadministrated DNase. Protection was 67%. Received data specify anopportunity and efficiency of DNase use for infections' treatment.

EXAMPLE 6 Level of Free Circulating Plasma DNA at Oncological Patientsand Healthy Adults

Blood plasma DNA of oncological patients and blood plasma DNA of healthydonors differs not only by its genetic repertoire but also by its amountcirculating in blood plasma. Table below presents data of blood plasmaDNA contents at 10 oncological patients and at 10 healthy volunteers.DNA isolation from plasma and determination of its amount was doneaccording to protocol mentioned above. DNA contents; Patient Sex, ageTumor, stage ng\ml 1 M, 67 Lung carcinoma 123 T2N2M0 2 F, 37 Breastcancer 78 T2N0M0 3 F, 53 Gaster carcinoma 90 T3N2M1 4 M, 54 Cancer oflarge intestine 340 T2N2M2 5 M, 64 Cancer of large intestine 182 T2N1M06 M, 56 Lung carcinoma 99 T3N2M1 7 F, 49 Cancer of large intestine 75T2N1M0 8 M, 65 Gaster carcinoma 120 T3N1M0 9 M, 36 Osteogenic sarcoma243 T3N1M2 10 F, 50 Breast cancer 165 T3N1M0 11 M, 24 Volunteer 10 12 M,32 Volunteer 27 13 F, 21 Volunteer 45 14 F, 19 Volunteer 7 15 F, 21Volunteer 13 16 F, 23 Volunteer 89 17 M, 28 Volunteer 11 18 F, 32Volunteer 15 19 F, 25 Volunteer 17 20 M, 38 Volunteer 5

As it is clear from the table the level of volunteers' blood plasma DNAis considerably lower than in blood of patients with different malignantneoplasms.

EXAMPLE 7 DNA Clones' Sequences, Received from Free Circulating BloodPlasma DNA of Patient with Malignant Mesothelioma

Clone 1

Duplicon, chromosome 15 and Y

Sequence No 1.

Clone 3

Unique, chromosome 2.

Sequence No 2

Clone 8

MLT2B repeat

Sequence No 3

Clone 9

Centromeric satellite DNA

Sequence No 4

Clone 10

MLT2B repeat

Sequence No 5

Clone 20

L1MC4-like (LINE-element)

Sequence No 6

Clone 15

Alpha-satellite DNA

Sequence No 7

Clones 18, 21

Alpha-satellite DNA

Sequence No 8

Clone 24

Unique, family of G protein-bound proteins, chromosome 6.

Sequence No 9

Clone 25

Unique, chromosome 3.

Sequence No 10

Clone 26

SatB1/Vimentin/nuclear matrix binding DNA

Sequence No 11

Sequence 33

Duplicon specific to the chromosome 10

Sequence No 12

Clone 32

alpha-satellite DNA

Clone 35

LTR repeat

Sequence No 13

Clone 36

Unique, chromosome 18

Sequence No 14

Clone 37

Unique, chromosome 4

Sequence No 15

Clone 41

Sequence No 16

Clone 43

Snf2-related CBP activator protein (SCRAP)

Unique, chromosome 16

Sequence No 17

Clone 45

Unique, chromosome 3

Sequence No 18

Clone 47

Alpha-satellite DNA

Clone 51

SRY-box containing gene.

Sequence No 19

Clone 52

Repeat

Sequence No 20

Clone 53, 55

Alpha-satellite DNA

Sequence No 21

Clone 56

Centromeric repeat

Sequence No 22

Clone 60

Gene repeated on several chromosomes, contains MER5A repeat.

Sequence No 23

Clone 62

Repeat

Sequence No 24

Clone 65

Unique, chromosome 2

Sequence No 25

Clone 71

Unique, chromosome 2

Sequence No 26

Clone 72

Unique, chromosome 8

Sequence No 27

Clone 73

Unique

Sequence No 28

Clone 78

Transposon Tigger fragment

Sequence No 29

Clone 81

Sequence No 30

Repeat (LINE)

Clone 82

Unique, chromosome 1

Sequence No 31

Clone 83

Unique, Fibroblast activation protein alpha; cell surface serineprotease

Chromosome 2

Sequence No 32

Clone 79

Alpha-satellite DNA

Clone 86

Unique, gene highly similar to brain testican, chromosome 4.

Sequence No 33

Clone 90

Unique, chromosome X

Sequence No 34

Clone 93

Unique, chromosome 9

Sequence No 35

Clones 89

92

Alpha-satellite DNA

Clone 96

Fragment LINE.

Sequence No 36

Clone 97

Chromosome 2 unique, Lipin

Clone 98

Unique, chromosome X

Sequence No 38

Clone 102

Chromosome 17 unique

Sequence No 39

Clone 99

Alpha-satellite DNA

Clone 105

Unique, chromosome 13

Sequence No 40

Clone N106

Chromosome 9 unique

Sequence No 41

Clone 107

Unique, chromosome 8

Sequence No 42

Clone N 111

Unique, chromosome 12

Sequence No 43

Clone N 112

Chromosome 5 unique

Sequence No 44

Clone 114

Chromosome 8 unique; Interleukin 7

Sequence No 45

Clone 116

Chromosome 1 unique

Sequence No 46

Clone 121

Chromosome 5 unique; Dynein

Sequence No 47

Clone 115; 119; 120

Alpha-satellite DNA

Clone 125

Chromosome 9 unique

Sequence No 48

Clone 127

Unique chromosome 20

Sequence No 49

Clone 130

Unique, chromosome is not determined.

Sequence No 50

Clone 124

SatB1/Vimentin/nuclear matrix binding DNA

Clone 133

Alpha-satellite DNA

Clone 137

MLT1A2 repeat

Sequence No 51

Clone 140

Unique, chromosome 2; zinc finger protein, subfamily 1A

Sequence No 52

Clone 141

Chromosome 2 unique

Sequence No 53

Clone 143

Fragment of Alu-repeat

Sequence No 54

Clone 144

Chromosome 2 unique

Sequence No 55

Clone 146

Chromosome 4 unique

Sequence No 56

Clone 139 and 142

Alpha-satellite DNA

Clone 148

Repeat (chromosomes 1, 2 and 4)

Sequence No 57

Clone 152

Unique, chromosome 16; KRAB-Domain, zinc finger protein

Sequence No 58

Clone 154

Chromosome 9 unique

Sequence No 59

Clone 161

Fragment LINE

Sequence No 60

Clone 151

Chromosome 5 unique

Sequence No 61

Clone 150

Chromosome 1 unique

Sequence No 62

Clone 153

Chromosome 11 unique

Sequence No 63

Clone 159

Chromosome 6 unique

Sequence No 64

Clone 163

Alpha satellite DNA

Sequence No 65

Clone 166

Chromosome 12 unique

Sequence No 66

Clone 167

Unique, chromosome 18, CDH2; cadherin 2, type 1, N-cadherin

Sequence No 67

Clones 169, 170

Chromosome 18 unique

Sequence No 68

Clone 178

Unique chromosome 1; RAMP: RA-regulated nuclear matrix-associatedprotein

Sequence No 69

Clone 180

Unique, chromosome 20

Sequence No 70

Clone 181

Unique chromosome 18

Sequence No 71

Clone 185

Alpha-satellite DNA

Sequence No 72

Clone 187

Mer repeat

Sequence No 73

Clone 188

HSATII repeat

Sequence No 74

Clone 189

Chromosome 9 unique

Sequence No 75

Clone 190

Chromosome 1 unique; melanoma antigen recognized by T cells 2

Sequence No 76

Clone 195

Chromosome 10 unique

Sequence No 77

Clone 196

Chromosome X unique

Sequence No 78

Clone 197

Chromosome 1 unique, FAF 1: Fas (TNFRSF6) associated factor 1

Sequence No 79

Clone 200

Chromosome 8 unique

Sequence No 80

Clone 202

Unique chromosome 13

Sequence No 81

Clone 205

Alpha satellite DNA

Sequence No 82

Clone 206

Repeat

Sequence No 83

Clone 208

Unique chromosome 8; Human DEAD box RNA helicase-like protein

Sequence No 84

EXAMPLE 8 DNA Clones Sequences Received From Free Circulating BloodPlasma DNA of Healthy Donor

Clone 1

Chromosome 5 unique

Sequence No 85

Clone 9

Unique chromosome 21

Sequence No 86

Clone 7

Unique chromosome 3

Sequence No 87

Clone 8

Chromosome 4 unique

Sequence No 88

Clone 10

18S RNA gene

Sequence No 89

Clone 11

Alu repeat

Sequence No 90

Clone 13

Unique chromosome 3

Sequence No 91

Clone 15

Unique chromosome 1

Sequence No 92

Clone 16

Unique chromosome 3, neutral endopeptidase

Sequence No 93

Clone 17

Chromosome 8 unique

Sequence No 94

Clone 18

Chromosome 1 unique

Sequence No 95

Clone 21

Unique chromosome 19; Zinc Finger protein

Sequence No 96

Clone 22

Unique chromosome 18

Sequence No 97

Clone 23

Unique chromosome 7, muskelin 1

Sequence No 98

Clone 25

Unique chromosome 11

Sequence No 99

Clone 27

Repeat

Sequence No 100

Clone 29

Unique chromosome 6

Sequence No 101

Clone 30

Unique chromosome 14

Sequence No 102

Clone 31

Unique chromosome 17

Sequence No 103

Clone 32

MER4B repeat

Sequence No 104

Clone 33

Chromosome 1 unique

Sequence 105

Clone 34

Unique chromosome 2

Sequence 106

Clone 35

Repeat

Sequence 107

Clone 36

Chromosome 1 unique

Sequence No 108

Clone 37

HERVH repeat

Sequence No 109

Clone 41

Chromosome X unique

Sequence No 110

Clone 42

Chromosome 6 unique

Sequence No 111

Clone 43

Unique chromosome 22; KREMEN1

Sequence No 112

Clone 44

Unique chromosome 14

Sequence No 113

Clone 45

Unique

Sequence No 114

Clone 46

Chromosome 20 unique

Sequence No 115

Clone 47

Nf-kappaB

Sequence No 116

Clone 38

Unique chromosome 16

Sequence No 117

Clone 48

Chromosome 6 unique

Sequence No 118

Clone 53

Unique

Sequence No 119

Clone 51

Chromosome 5 que

Sequence No 120

Clone 59

Unique chromosome 4, NFKB 1: nuclear factor of kappa light polypeptidegene enhancer

Sequence No 121

Clone 61

Repeat

Sequence No 122

Clone 62

L1 repeat

Sequence No 123

Clone 64

Duplicon chromosome 7

Sequence No 124

Clone 65

Ribosomal DNA

Sequence No 125

Clone 66

Rbosomal DNA

Sequence No 126

Clone 75

Repeat

Sequence No 127

Clone 76

Chromosome 4 unique

Sequence No 128

Clone 83

Chromosome 4 unique

Sequence No 129

Clone 85

Unique chromosome 2; phospholipase C, epsilon

Sequence No 130

Clone 87

L1PA3 repeat

Sequence No 131

Clone 86

Unique chromosome 5; CRTL 1: cartilage linking protein 1

Sequence No 132

Clone 89

Alu repeat

Sequence No 133

OH 92

Unique chromosome 6

Sequence No 134

Clone 100

Unique, chromosome 6

Sequence No 135

Clone 105

AluSx repeat

Sequence No 136

Clone 111

Alphoid repetitive DNA

Sequence No 137

Clone 112

Chromosome 9 unique

Sequence No 138

Clone 113

Chromosome 22 unique

Sequence No 139

Clone 114

AluSx repeat

Sequence No 140

Clone 116

Unique chromosome 9; 17 kD fetal brain protein

Sequence No 141

Clone 123

Unique chromosome 5

Sequence No 142

Clone 124

Unique chromosome 13

Sequence No 143

Clone 126

Unique chromosome 8

Sequence No 144

Clone 130

Unique chromosome 1

Sequence No 145

Clone 131

Unique chromosome 4

Sequence No 146

Clone 136

Unique chromosome 8

Sequence No 147

Clone 141

Unique chromosome 2

Sequence No 148

Clone 146

Unique chromosome 16

Sequence No 149

Clone 147

Unique chromosome 5; nicotinamide nucleotide transhydrogenase

Sequence No 150

Clone 149

Unique chromosome 9

Sequence No 151

Clone 151

Unique chromosome 16

Sequence No 152

Clone 152

Unique chromosome 6, BAI3: brain-specific angiogenesis inhibitor 3

Sequence No 153

Clone 153

Unique chromosome 9, GAD2: glutamate decarboxylase 2

Sequence No 154

Clone 155

Unique chromosome 9

Sequence No 155

EXAMPLE 9 Sensitivity of Circulating Blood DNA to DNase Action

DNA from fresh blood serum (mixture of blood received from 5 volunteers) was isolated by standard phenol-chloroform method.

Sediment of nucleic acids was washed by 70% ethanol and dissolved intris-EDTA buffer. Spectrophotometry with wave's length 260 nm (SF-46)was used for quantification of DNA. DNA was researched byelectrophoresis in 0.8% polyacrylamide gel with ethidium bromidestaining. One part of received DNA was processed by DNase I for 3 hoursat 37° C. After electrophoretic separation of processed and notprocessed DNA it was found out that:

After electrophoresis non-processed DNA constitute one compact band.This factor indicates on relatively identical size of DNA circulating inblood. Said band disappeared completely after DNAse processing. Receiveddata indicates that DNA circulating in serum is sensitive to DNases useand can be destroyed by their action.

EXAMPLE 10 Results of the Experiment of Influence of Polyclonal SerumContaining Anti-DNA Antibodies' on Life Span of Erlich Carcinoma BearingMice

DNA circulating in blood plasma of oncological patients can be destroyedor inactivated not only by DNA—digesting enzymes (for example by DNase),but also by other methods.

Anti-DNA antibodies were isolated from blood of patients with lupuserythematosus by Shuster A. M. method (Shuster A. M. et. al., Science,v. 256, 1992, pp. 665-667). Such anti-DNA antibodies were capable notonly to bind DNA but also to hydrolyze it.

Group 1—7 mice transplanted with Erlich carcinoma (control).

Group 2—6 mice transplanted withy Erlich carcinoma. Mice were injectedintravenously with fraction of human anti-DNA antibodies (Ig G) at 200mkg dose on the third day after tumor transplantation.

Group 3—6 mice transplanted withy Erlich carcinoma. Mice were injectedintravenously with fraction of human non specific immunoglobulin (Ig G)at 200 mkg dose on the third day after tumor transplantation.

Effect was estimated according to tumor's growth inhibition on the7^(th) day after transplantation (TGO expressed in percents).

Tumor's on the 7^(th) day after transplantation. Group Tumor's volume T%P 1  85 +/− 12 — — 2 45 +/− 6 52% p < 0.001 3 79 +/− 7  7% p < 0.001

Results of experiments indicates that single anti-DNA antibodies'administration possess strong antitumor effect. Administration ofantibodies fraction that does not contain ant-DNA antibodies do notpossess antitumor activity.

EXAMPLE 11 Effect of Mice' Vaccination by Blood Plasma DNA FractionReceived from Animals with Erlich Carcinoma on Transplantation andGrowth of Erlich Carcinoma at Immunized Animals

DNA from plasma of mice with Erlich carcinoma was isolated on the 5thday after transplantation according to method mentioned above.Positively charged multilayer liposomes were used as adjuvants forimmunization. DNA sample was mixed with liposomes (20 mkg of DNA in 1 mgof lipids).

Group 1—6 non immunized mice.

Group 2—6 mice immunized for three times with one week 5 interval byintradermal administration of 1 mkg DNA from blood plasma in 50 mkl ofliposomal suspension containing 50 mkg of liposomes.

Group 3—6 mice immunized for three times with one week interval byintradermal administration of 50 mkl of liposomal suspension containing50 mkg of liposomes without DNA.

Group 4—6 mice immunized for three times with one week interval byintradermal administration of 1 mkg DNA from calf thymus (Sigma) in 50mkl of liposomal suspension containing 50 mkg of liposomes.

One week after the last immunization Erlich carcinoma was transplantedto all mice including non immunized control mice.

Results of the experiment were estimated according to animal survival onthe 30th and 50^(th) day after tumor's transplantation. 30 day 50 day(number of alive\ (number of alive\ number of dead in number of dead inGroup group) group) 1 0-6 0-6 2 5-6 3-6 3 0-6 0-6 4 2-6 0-6

So immunization of mice with replanted Erlich carcinoma by blood plasmaDNA lead to significant increasing of animals' lifetime.

EXAMPLE 12 DNA Isolation from Biofilm Matrix Formed by Gram-Positive andGram-Negative Bacteria

Biofilms of Escherichia coli and Staphylococcus aureus were used inexperiments. Biofilms were washed off by phosphate buffer from agar.Cells and matrix were separated by centrifuging. Standardphenol-chloroform method was used for DNA extraction from matrix. DNAamount was estimated by spectrophotometric analysis with wave's length260 nm (SF-46). Received DNA was researched by electrophoresis in 0.8%polyacrylamide gel with ethidium bromide staining.

One part of received DNA was processed for 3 hours by DNAase I at 37° C.After electrophoretic separation of processed and not processed DNA itwas found out that:

After electrophoresis non-processed DNA form one compact line. This factindicates relatively identical size of DNA in biofilm matrix. Said banddisappeared completely after DNAse processing. So the biofilm matrix ofgram positive and gram negative bacteria contains extracellular DNA,which could be destroyed with DNAse.

EXAMPLE 13 Dynamics of P-Glycoprotein Expression in Erlich Carcinoma atMice that Receive Doxorubicin Therapy and DNase I Effect

Treatment by Doxorubicin causes expression of P-glycoprotein in tumortissue that is one of the main MDR (Multidrug Resistance) phenotypemediators. Immunohistochemical staining of mice's tumor histologicalcuts are listed below.

Mice were subjected to course of 5 day therapy with Doxorubicin (2 mg/kgintravenously daily) or Doxorubicin+DNase I (0.5 mg/kg four times a dayduring 5 days)

Treatment has begun on the 3^(rd) day after tumor's transplantation.Tissue preparations were executed on the 8th day of tumor'stransplantation. Multifocal expression of P-glycoprotein was observed intumor tissue after 5 days of therapy (FIG. 1).

Total level of P-glycoprotein expression and amount of P-glycoproteinpositive nodules in tumor tissue was much lower at the case of combinedtreatment by Doxorubicin+DNase (FIG. 1). So treatment by DNase delaysdevelopment of multidrug resistant phenotype in tumor that is caused byantitumor antibiotic Doxorubicin use.

EXAMPLE 14 Influence of Plasma DNA of C57B1 Mice with LLC Tumor AfterChemotherapeutical Treatment by Doxorubicin, on LLC Tumor Growth andMetastasizes Development at C57B1 Mice Receiving Doxorubicin Therapy andEffect of DNase I

LLC tumor was replanted to 30 C57B1 mice. Twenty mice were treated withDoxorubicin at 2 mg/kg dose daily for 5 days, starting day 3 aftertransplantation. Ten mice were treated with Cyclophosphamide at 15 mg/kgdose intraperitoneally for once on the 3^(rd) day after replantation.Such treatment scheme does not lead to animal's recovery but leads to50% tumor inhibition at day 8 in doxorubicin treated animals and 30%tumor inhibition at day 8 in Cyclophosphamide treated animals. On thenext day after end of chemotherapy course animals were euthanized andand total blood plasma from both mice groups was taken. After isolationtotal fraction of blood plasma DNA was stored at −20° C. in phosphatebuffer.

Five groups of mice that were transplanted with LLC tumor participatedin experiment.

Group 1—7 mice (control).

Group 2—6 mice intravenously treated with Doxorubicin chemotherapy from3^(rd) up to 8^(th) day at 2 mg/kg dose daily.

Group 3—6 mice intravenously treated with Doxorubicin chemotherapy from3^(rd) up to 8^(th) day at 2 mg/kg dose daily+intravenous administrationof DNA fraction from mice previously subjected to Doxorubicinchemotherapy (0.05 mkg of DNA in 200 mkl of phosphate buffer at day 1and day 3 after initiation of treatment)

Group 4-6 mice intravenously treated with Doxorubicin chemotherapy from3^(rd) up to 8th day at 2 mg/kg dose daily+intravenous administration ofDNA fraction from mice previously subjected to cyclophosphamidechemotherapy (0.05 mkg of DNA in 200 mkl of phosphate buffer at day 1and day 3 after initiation of treatment)

Group 5—6 mice intravenously treated with Doxorubicin chemotherapy from3^(rd) up to 8^(th) day at 2 mg/kg dose daily+intravenous administrationof DNA fraction from mice previously subjected to Doxorubicinchemotherapy (0.05 mkg of DNA in 200 mkl of phosphate buffer at day 1and day 3 after initiation of treatment)+intraperitoneal administrationof DNase I at 0.5 mg/kg dose for 4 times a day at the first and seconddays of treatment.

Tumor's size on the 8^(th) day after transplantation. Group Tumor's size1 127+/−13 2 67 +/− 7 3 115 +/− 20 4  75 +/− 11 5 82 +/− 9

So administration of blood plasma DNA from mice subjected tochemotherapy lead to tumor's resistance to chemotherapeutic treatment.DNase's administration prevents appearance of this effect.

EXAMPLE 15 Influence of Blood Plasma DNA from C57B1 Mice with HighlyMetastatic LLC Strain on Metastasizing of Low Metastatic LLC TumorStrain in C57B1 Mice and Effect of DNase I

LLC tumor was transplanted to 30 C57B1 mice. Twenty mice weretransplanted with highly metastatic strain and 10 mice were transplantedwith low metastatic strain. On the 9th day animals were euthanized andtotal blood plasma of both mice groups was collected. After isolationthe total fraction of blood plasma DNA was stored at −20° C. inphosphate buffer.

Five groups of mice with transplanted LLC tumor participated in theexperiment.

1 Group—6 mice transplanted with low metastatic LLC strain.

2 Group—6 mice transplanted with low metastatic LLC strain+intravenousadministration of total DNA fraction from mice with transplanted highlymetastatic strain (0.05 mkg of DNA in 200 mkl of phosphate buffer on the7^(th) and 8^(th) day after transplantation).

3 Group—6 mice transplanted with low metastatic LLC strain+intravenousadministration of total DNA fraction from mice with transplanted lowmetastatic strain (0.05 mkg of DNA in 200 mkl of phosphate buffer on the7^(th) and 8^(th) day after transplantation)

4 Group—6 mice transplanted with low metastatic LLC strain+intravenousadministration of total DNA fraction from mice with transplanted highlymetastatic strain (0.05 mkg of DNA in 200 mkl of phosphate buffer on the7^(th) and 8^(th) day after transplantation)+intraperitonealadministration of DNase I at 1 mg/kg dose two times daily at 7^(th) and8^(th) day after transplantation.

5 Group—6 mice transplanted with highly metastatic LLC strain.

Number of metastatic foci in lungs was estimated bon the 15^(th) dayafter transplantation (N).

Experiments' results are presented in the table. Group N 1 12, 0 2 24, 13 14, 6 4 11, 6 5 33, 6

Received data indicates that blood plasma DNA from mice with highlymetastatic LLC strain intensify metastasizing of low metastatic LLCstrain.

DNase administration prevents appearance of this effect.

EXAMPLE 16 DNase I Influence on Life Span of C57B1 Mice Transplantedwith LLC Tumor (Highly Metastatic Strain)

Five groups of LLC transplanted mice participated in the experiment.

Group 1—7 mice (control).

Group 2—6 mice were treated with intraperitoneal administration of DNaseat 1 mg/kg dose two times a day starting from 3 up to 7 day after tumortransplantation.

Group 3—6 mice were treated with intraperitoneal administration of DNaseat 1 mg/kg dose two times a day starting from 3 up to 10 day after tumortransplantation.

Group 4—6 mice were treated with intraperitoneal administration of DNaseat 1 mg/kg dose two times a day starting from 3 up to 15 day after tumortransplantation.

Group 5—6 mice were treated with intraperitoneal administration of DNaseat 1 mg/kg dose two times a day starting from 3 up to 18 day after tumortransplantation.

Results of experiment were estimated according to animals' survival onthe 30 and 50 day after tumor transplantation. 30 day 50 day (number ofalive\ (number of alive\ number of dead in number of dead in Groupgroup) group) 1 0-7 0-7 2 0-6 0-6 3 3-6 0-6 4 5-1 3-3 5 6-0 6-0

The significant inhibition of tumor's growth was observed at the lastday of DNase treatment in the 2nd and 3rd groups, but tumor's growthrenewed after DNase withdrawal and to the 25^(th) day size of tumor inthis groups and in control has equalized.

The most longitudinal course o DNase treatment (from 3^(rd) up to 18thday—group number 6) has lead to maximal survival. Inhibition of tumorgrowth was more than 95% at day 18.

In all experiments single and multiple injection of up to 2.5 mg/kg ofhuman DNase I (maximal dose that was used in experiments) had no toxiceffect on animals.

So, DNase I does not cause direct cytotoxic effect on tumor cells (inour in vitro experiments at concentration of 100 mkg/ml) andexperimental data confirm that antitumor effect is connected withdestruction of DNA in blood plasma and DNase's therapeutic effectincreases with increasing of its treatment course duration.

EXAMPLE 17 Influence of Different Methods of Blood Plasma DNA'sDestruction, Inactivation, and Binding on Ability of Blood Plasma DNAfrom C57B1 Mice with Transplanted Highly Metastatic LLC Strain toIntensify Metastasizing of Low Metastatic LLC Tumor Strain in C57B1 Mice

100 mice were transplanted with highly metastatic LLC strain. On the 9thday after transplantation, animals were euthanized and total bloodplasma was taken. After isolation total fraction of blood plasma DNA wasstored at −20° C. in phosphate buffer.

Six groups of mice with transplanted low metastatic LLC strainparticipated in the experiment.

Group 1—6 mice transplanted with low metastatic LLC strain.

Group 2—6 mice transplanted with low metastatic LLC strain+twointravenous injections of total DNA fraction from mice transplanted withhighly metastatic strain on the 7^(th) and 8^(th) day aftertransplantation (0.05 mkg of DNA was dissolved in 500 mkl of freshheparinized blood before injection).

Group 3—6 mice transplanted with low metastatic LLC strain+twointravenous injections of total DNA fraction from mice transplanted withhighly metastatic strain on the 7^(th) and 8^(th) day aftertransplantation (0.05 mkg of DNA was dissolved in 500 mkl of fresh bloodplasma before injection). Before administration the sample wasphotochemically disinfected (1 mkM of methylene blue was added withfollowing irradiation by red light during 10 minutes (60 000 Lux).

Group 4—6 mice transplanted with low metastatic LLC strain+twointravenous injections of total DNA fraction from mice transplanted withhighly metastatic strain on the 7^(th) and 8^(th) day aftertransplantation (0.05 mkg of DNA was dissolved in 500 mkl of fresh bloodplasma before injection). The sample was passed through the columncontaining DEAE-cellulose for two times before administration.

Group 5—6 mice transplanted with low metastatic LLC strain+twointravenous injections of total DNA fraction from mice transplanted withhighly metastatic strain on the 7^(th) and 8^(th) day aftertransplantation (0.05 mkg of DNA was dissolved in 500 mkl of freshheparinized blood before injection. 1 mkg of fragment A of Ricin toxinwas added to the sample before administration and sample was incubatedat 370C for 1 hour. Ricin toxin is representative of RIP toxins family(proteins that inactivate ribosomes) which are used for immunotoxin'screation. Besides their ability to inactivate ribosomes these proteinscan deadenylate DNA. For realization of toxic effect catalytic subunit Aof RIP II type should by delivered to cell by B subunit. Without Bsubunit A chain is not toxic but can be used for blood plasma DNA'sinactivation due to its polynucleotide-adenylglicozidase activity.

Group 6—6 mice transplanted with low metastatic LLC strain+twointravenous injections of total DNA fraction from mice transplanted withhighly metastatic strain on the 7^(th) and 8^(th) day aftertransplantation (0.05 mkg of DNA was dissolved in 500 mkl of freshheparinized blood before injection. Total DNA fraction was enzymaticallymethylated before administration (I. Muiznieks et. al., FEBS Letters,1994, v. 344, pp. 251-254).

Number of metastaic nodules in lungs was estimated on the 15th day aftertransplantation.

Results of the experiments are presented in the table. Group Ncp. 1 12,0 2 22, 5 3 14, 1 4 15, 5 5 15, 1 6 12, 3

Received data indicates that all used methods inhibited ability of bloodplasma DNA of mice with highly metastatic LLC tumor strain to increasemetastasizing process of low metastatic LLC tumor strain.

EXAMPLE 18 Influence of DNase I Therapy on the Development of SomaticMosaicism

Frequency of HPRT gene's mutations in blood T-lymphocytes was studied asa model of somatic mosaicism. Human HPRT gene (chromosome Xq26) encodesenzyme that is constitutionally expressing but not essential and isinvolved in metabolism of purine nucleotides. Cloning was done accordingto Bigbee W. method (Bigbee W. Et al., Mutation Res., 1998, v. 397, pp.119-136).

Lymphocytes from peripheral blood of 8 patients subjected to 3 weekcourse of immunostimulating therapy by Neovir after surgical eliminationof tumor were used for cloning. 4 patients out from 8 were additionallysubjected to intravenous administration of human recombinant DNase I(200 mkg/kg dose 4 times a day during three weeks).

Incidence of HPRT—deficient clones in the blood of patients thatreceived DNase I therapy was 3 times less than in blood of patients thatreceived only immunostimulating therapy.

EXAMPLE 19 Influence of DNase I Therapy on Old Rats' Life Time

White not thoroughbred 24 month old rats were used in experiment. In thestudy group (10 animals) rats starting from the age of 24 months weresubjected to intravenous administration of polysialated human DNase I at500 mg/kg (equivalent dose of non-modified enzyme) 2 times a week during2 months. Control group of animals got the administration of placebo.Average life time of rats from control group was 27.8 months. Averagelife time of rats from experimental group was 30.1 months.

EXAMPLE 20 Influence of DNase I Therapy on Viability of Pancreatic{tilde over (β)} Cells and Endothelium of Aorta

{tilde over (β)} cells of human embryonic pancreas and endothelial cellsof human aorta were used for primary cell culture formation. DNAisolated from plasma of patient with severe II-type diabetes complicatedby systemic atherosclerosis (0.0025 mkg per 1 ml of culture media) wasadded to cell culture 24 hours after passage.

In other experimental series DNA of the patient before introduction tocell culture was processed with enzymatic methylation. Amount of viablecells was estimated 24 hours later with trypan blue staining.

Results of experiment are presented in the table.

Percentage of viable cells 48 hours after subculturing. Cells ControlPatient's DNA Methylated DNA B-cells 73% 43% 61% Endothelium 62% 35% 55%

INDUSTRIAL APPLICABILITY

Current invention justify that destruction (binding, inactivation) ofcirculating blood plasma DNA in course of oncological and bacterialdiseases possess significant therapeutic effect.

Invention is confirmed by the fact that blood plasma DNA of patientscontains unique genes that takes part in formation and maintenance oftumor's “malignant phenotype”.

Binding, destruction or inactivation of DNA circulating in blood plasmapossess therapeutic effect at diseases which development is connectedwith accumulation and spreading of somatic mutations in host's cells.

Blood plasma circulating DNA can be destructed, inactivated oreliminated from blood plasma by DNases, sorbents, antibodies and othermethods which provide inactivation by destroying, binding ormodification of circulating DNA.

Treatment directed to the destruction of blood plasma DNA lead tosignificant antitumor effect while its own toxicity is immaterial.

Treatment directed to the destruction of blood plasma DNA if combinedwith traditional antitumor therapy lead to significant antitumor effect.

Efficiency of the treatment directed to blood plasma DNA's destructioncan be estimated by monitoring of DNA amount and amount of tumor'sgenetic markers in blood plasma of patient who receive such therapy.

Cloning and sequencing of blood plasma DNA from oncological patient'scan be used for studying of genetic processes that take part inoncogenesis and identification of new genes connected with oncogenesisprocesses.

Cloning and sequencing of healthy people's blood plasma DNA can be usedfor studying of genome's operating in healthy individual and in thedevelopment of somatic diseases and in identification of new genes thatare involved in these processes.

Pharmaceutical compositions containing components that inactivate bydestroying, binding and modification the blood plasma DNA could be usedfor treatment of patients with oncological diseases, infections, somaticdiseases and for prolongation of life time.

For providing effective therapeutic exposition of active component thatis necessary for blood plasma DNA' destruction, and for therapeuticeffect achievement pharmaceutically acceptable compositions andmodifications, drug delivery systems are used providing maximal systemiccirculations of the active component in blood plasma and its contactwith DNA circulating in plasma. The main way of administration isintravenous. But other ways of administration that provide coming ofactive component in to systemic circulation, such as subcutaneous,intramuscular, inhalation, intraperitoneal and others can also be used.Doses and regimes of administration are determined by nature of usedactive ingredient and way of administration. The effect is controlled bythe level of DNA's contents and its dynamics in blood plasma, presenceof tumor, infectious and other genetic markers, and appearing ofpositive clinical dynamics of disease.

1. Treatment method of oncological and/or infectious and/or somatic diseases by acting on biological targets inside organisms, differs in that the biological target is extracellular DNA including extracellular DNA circulating in blood plasma.
 2. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1 differs in that the extracellular DNA is inactivated by destruction, binding or modification.
 3. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1 and 2, differs in that the extracellular DNA is inactivated by destruction, binding or modification by injecting to patient of pharmaceutical agent which is capable to destroy, bind or modify free circulating DNA.
 4. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1-3, differs in that the extracellular DNA is inactivated by destruction, binding or modification by pharmaceutical agent's injection in amount sufficient for destruction and in therapeutic regime providing destruction, binding or modification in sufficient for therapeutic effect achievement period of time.
 5. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1-4 differs in that the genetically modified cells or genotherapeutic constructions are injected to patient when said remedies induce synthesis in host's organism of biopolymers, capable to inactivate free circulating blood plasma DNA by its binding, destruction or modification.
 6. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1 or 2, differs in that the circulating extracellular DNA is inactivated by destruction, binding or modification using extracorporeal blood processing.
 7. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1, 2 or 6, differs in that the extracorporal purification of patient's blood from free circulating DNA is achieved by immune or affine absorption.
 8. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1, 2 or 6, differs in that the extracorporal purification of patient's blood from free circulating DNA is achieved by methods of gravitational blood surgery.
 9. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1, 2 or 6, differs in that the extracorporal purification of patient's blood from free circulating DNA is achieved by biological, chemical or photochemical inactivation.
 10. Treatment method of oncological and/or infectious and/or somatic diseases according claim 1 or 2, differs in that the patient is immunized by vaccine, which vaccine contain blood plasma circulating DNA (including said DNA with naturally complexed proteins) as the antigen.
 11. Treatment method of oncological and/or infectious and/or somatic diseases according claims 1-10, differs in that the treatment is combined with surgical, chemiotherapeutic, hormonal, radiation and immunotherapeutic methods.
 12. Pharmaceutical agent for oncological and/or infectious and/or somatic disease treatment, representing compound possessing desoxyribonuclease activity and/or being able to inactivate extracellular DNA including DNA circulating in patients blood plasma.
 13. Pharmaceutical agent according claim 12 differs in that compound possessing desoxyribonuclease activity is desoxyribonuclease enzyme.
 14. Pharmaceutical agent according claim 13 differs in that desoxyribonuclease is modified for better pharmacodynamic and pharmacokinetic performance and comprises desoxyribonuclease analogue with increased activity, desoxyribonuclease analogue not sensitive to endogenous inhibitors of desoxyribonuclease, polysialated desoxyribonuclease, pegylated desoxyribonuclease, desoxyribonuclease that is bound or mixed with synthetic and natural microspheres, liposomes, dextran, and other corpuscular natural and synthetic polymer carriers.
 15. Pharmaceutical agent according claims 12-14 which additionally contains ribonuclease and/or lipase and/or proteinase.
 16. Pharmaceutical agent according claim 12, differs in that the compound possessing desoxyribonuclease activity is antibody possessing nuclease activity, in particular polyclonal DNA-abzymes, monoclonal DNA-abzymes or their derivatives.
 17. Pharmaceutical agent according claim 12, differs in that the compound able to bind DNA is antibody able to bind DNA and its complexes and derivatives of said antibody.
 18. Pharmaceutical composition for oncological and infectious diseases treatment, containing pharmaceutical agent according claims 12-16 in therapeutically effective amount and pharmaceutically acceptable carrier or excipient.
 19. Method to increase the life time which is achieved by inactivation of extracellular DNA circulating in blood plasma by said DNA destruction, binding or modification according claims 2-17.
 20. Method of prophylaxis of pathologies connected with appearance and development of somatic mosaicism by the way of destruction, binding or modification of DNA according to claims 2-17.
 21. Method to control the treatment efficacy of oncological and/or infectious and somatic diseases, to estimate the infection development, to control the efficacy of treatment directed to prolongation of life time, by the way of measurement of patient biochemical factors, differs in that monitoring for control of such treatment sizes of molecules, fractions' correlation, bindings with proteins, lipids and sugars, nucleotide consequences of free circulating blood plasma DNA are used.
 22. Usage of blood plasma DNA and extracellular microbial DNA for evaluation of DNA involved in process of diseases' appearance and development, which usage includes its cloning, sequencing, identification of genes, unique and repeated sequences with their future studying. 